ELISA Kit for Estradiol (E2)

17B-Estradiol; Oestradiol; Beta-Estradiol


This assay has high sensitivity and excellent specificity for detection of Estradiol (E2).
No significant cross-reactivity or interference between Estradiol (E2) and analogues was observed.


Matrices listed below were spiked with certain level of Estradiol (E2) and the recovery rates were calculated by comparing the measured value to the expected amount of Estradiol (E2) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 85-92 88
EDTA plasma(n=5) 80-101 88
heparin plasma(n=5) 96-104 101


Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Estradiol (E2) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Estradiol (E2) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%


The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Estradiol (E2) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 89-104% 83-102% 86-101% 80-95%
EDTA plasma(n=5) 81-96% 83-94% 80-95% 78-101%
heparin plasma(n=5) 86-101% 80-94% 80-101% 84-101%


The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.



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Stem Cell Research & Therapy G-CSF-mobilized Peripheral Blood Mononuclear Cells Combined with Platelet-rich Plasma Restored the Ovarian Function of aged rats via angiogenesis and …
Research Square Drug-free in vitro activation combined 3D-bioprinted adipose-derived stem cells restore ovarian function of premature ovarian insufficiency
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